Inhibition of Transforming Growth Factor-Activated Kinase 1 (TAK1) Blocks and Reverses Epithelial to Mesenchymal Transition of Mesothelial Cells
Figure 1
TAK1 inhibition leads to the reversal of the EMT phenotype (MET) in MCs derived from peritoneal effluent of PD patients.
A, Confocal immunofluorescence analysis (red) of MCs from a patient undergoing PD. Cells were treated with DMSO, NP-009245, (600 nM), CI 1040 (2 µM) for 72 h. Cells were fixed and stained with rhodamine phalloidin. Nuclei were stained with Hoechst 33342 (blue). The immunofluorescence shown is representative of three independent experiments. B, Western blots showing the expression of E-cadherin in total cell lysates of MCs. Cells were treated with DMSO, NP-009245 (T) (600 nM), CI 1040 (CI) (2 µM), or a combination of the two pharmacological inhibitors for 72 h. Expression of tubulin was detected as a loading control. C, Effect of TAK1 pharmacological inhibition on E-cadherin mRNA expression in MCs from patients undergoing PD. Cells were treated as in A. Quantitative RT-PCR was performed on total RNA. Histone H3 mRNA levels were used for normalization. Bars represent means+s. e. m. of duplicate determinations in three independent experiments. D, Effect of TAK1 pharmacological inhibition on N-cadherin mRNA expression in MCs from patients undergoing PD. Cells were treated with DMSO or NP-009245, (600 nM) for 72 h. Quantitative RT-PCR was performed on total RNA. Histone H3 mRNA levels were used for normalization. Bars represent means+s. e. m. of duplicate determinations in three independent experiments. E and F, Confocal immunofluorescence analysis of MCs from patients undergoing PD. Cells were treated with DMSO or NP-009245, (600 nM) for 72 h. Cells were fixed and stained with a monoclonal antibody against cytokeratin (red) (D) or a polyclonal antibody against ZO-1 (green) (E). Nuclei were stained with Hoechst 33342 (blue). All data are representative of at least three independent experiments. * p<0.05 versus DMSO treated cells.